Journal Article

A cytosine deaminase for programmable single-base RNA editing

Koob, Jeremy, Kellner, Max J., Franklin, Brian, Joung, Julia, Abudayyeh, Omar O., Ladha, Alim, Kirchgatterer, Paul, Gootenberg, Jonathan S., Cox, David B. T., Zhang, Feng

2019

Overview

Programmable RNA editing enables reversible recoding of RNA information for research and disease treatment. Previously, we developed a programmable adenosine-to-inosine (A-to-I) RNA editing approach by fusing catalytically inactivate RNA-targeting CRISPR-Cas13 (dCas13) with the adenine deaminase domain of ADAR2. Here, we report a cytidine-to-uridine (C-to-U) RNA editor, referred to as RNA Editing for Specific C-to-U Exchange (RESCUE), by directly evolving ADAR2 into a cytidine deaminase. RESCUE doubles the number of mutations targetable by RNA editing and enables modulation of phosphosignaling-relevant residues. We apply RESCUE to drive β-catenin activation and cellular growth. Furthermore, RESCUE retains A-to-I editing activity, enabling multiplexed C-to-U and A-to-I editing through the use of tailored guide RNAs.

• Publisher: American Association for the Advancement of Science,The American Association for the Advancement of Science
• Subject: Adenine / Adenine deaminase / Adenosine / Adenosine - genetics / Adenosine Deaminase - chemistry / Adenosine Deaminase - genetics / beta Catenin - chemistry / beta Catenin - genetics / beta Catenin - metabolism / Biological evolution / CRISPR / Cytidine - genetics / Cytidine deaminase / Cytosine / Cytosine deaminase / Cytosine Deaminase - chemistry / Cytosine Deaminase - genetics / Deamination / Directed evolution / Editing / Exchanging / HEK293 Cells / Humans / Inosine - genetics / Medical treatment / Modulation / Mutation / Phosphorylation / Protein Domains / Protein Engineering - methods / RNA Editing / RNA-Binding Proteins - chemistry / RNA-Binding Proteins - genetics / Uridine / Uridine - genetics / β-Catenin